Rostral-caudal Axis Research Articles

Predicting neuronal response properties from hemodynamic responses in the auditory cortex

Recent functional MRI (fMRI) studies have highlighted differences in responses to natural sounds along the rostral-caudal axis of the human superior temporal gyrus. However, due to the indirect nature of the fMRI signal, it has been challenging to relate these fMRI observations to actual neuronal response properties. To bridge this gap, we present a forward model of the fMRI responses to natural sounds combining a neuronal model of the auditory cortex with physiological modeling of the hemodynamic BOLD response. Neuronal responses are modeled with a dynamic recurrent firing rate model, reflecting the tonotopic, hierarchical processing in the auditory cortex along with the spectro-temporal tradeoff in the rostral-caudal axis of its belt areas. To link modeled neuronal response properties with human fMRI data in the auditory belt regions, we generated a space of neuronal models, which differed parametrically in spectral and temporal specificity of neuronal responses. Then, we obtained predictions of fMRI responses through a biophysical model of the hemodynamic BOLD response (P-DCM). Using Bayesian model comparison, our results showed that the hemodynamic BOLD responses of the caudal belt regions in the human auditory cortex were best explained by modeling faster temporal dynamics and broader spectral tuning of neuronal populations, while rostral belt regions were best explained through fine spectral tuning combined with slower temporal dynamics. These results support the hypotheses of complementary neural information processing along the rostral-caudal axis of the human superior temporal gyrus.

Phase I double-blind, placebo-controlled trial of dolcanatide (SP-333) 27 mg to explore colorectal bioactivity in healthy volunteers

ABSTRACT Guanylyl cyclase C (GUCY2C) is a tumor-suppressing receptor silenced by loss of expression of the luminocrine hormones guanylin and uroguanylin early in colorectal carcinogenesis. This observation suggests oral replacement with a GUCY2C agonist may be an effective targeted chemoprevention agent. Previous studies revealed that linaclotide, an oral GUCY2C agonist formulated for gastric release, did not persist to activate guanylyl cyclase signaling in the distal rectum. Dolcanatide is an investigational oral uroguanylin analog, substituted with select D amino acids, for enhanced stability and extended persistence to activate GUCY2C in small and large intestine. However, the ability of oral dolcanatide to induce a pharmacodynamic (PD) response by activating GUCY2C in epithelial cells of the colorectum in humans remains undefined. Here, we demonstrate that administration of oral dolcanatide 27 mg daily for 7 d to healthy volunteers did not activate GUCY2C, quantified as accumulation of its product cyclic GMP, in epithelial cells of the distal rectum. These data reveal that the enhanced stability of dolcanatide, with persistence along the rostral-caudal axis of the small and large intestine, is inadequate to regulate GUCY2C across the colorectum to prevent tumorigenesis. These results highlight the importance of developing a GUCY2C agonist for cancer prevention formulated for release and activity targeted to the colorectum.

Global feather orientations changed by electric current

SummaryDuring chicken skin development, each feather bud exhibits its own polarity, but a population of buds organizes with a collective global orientation. We used embryonic dorsal skin, with buds aligned parallel to the rostral-caudal body axis, to explore whether exogenous electric fields affect feather polarity. Interestingly, brief exogenous current exposure prior to visible bud formation later altered bud orientations. Applying electric pulses perpendicular to the body rostral-caudal axis realigned bud growth in a collective swirl, resembling an electric field pointing toward the anode. Perturbed buds show normal molecular expression and morphogenesis except for their altered orientation. Epithelial-mesenchymal recombination demonstrates the effects of exogenous electric fields are mediated through the epithelium. Small-molecule channel inhibitor screens show Ca2+ channels and PI3 Kinase are involved in controlling feather bud polarity. This work reveals the importance of bioelectricity in organ development and regeneration and provides an explant culture platform for experimentation.

Cannabidiol has a unique effect on global brain activity: a pharmacological, functional MRI study in awake mice

BackgroundThe phytocannabinoid cannabidiol (CBD) exhibits anxiolytic activity and has been promoted as a potential treatment for post-traumatic stress disorders. How does CBD interact with the brain to alter behavior? We hypothesized that CBD would produce a dose-dependent reduction in brain activity and functional coupling in neural circuitry associated with fear and defense.MethodsDuring the scanning session awake mice were given vehicle or CBD (3, 10, or 30 mg/kg I.P.) and imaged for 10 min post treatment. Mice were also treated with the 10 mg/kg dose of CBD and imaged 1 h later for resting state BOLD functional connectivity (rsFC). Imaging data were registered to a 3D MRI mouse atlas providing site-specific information on 138 different brain areas. Blood samples were collected for CBD measurements.ResultsCBD produced a dose-dependent polarization of activation along the rostral-caudal axis of the brain. The olfactory bulb and prefrontal cortex showed an increase in positive BOLD whereas the brainstem and cerebellum showed a decrease in BOLD signal. This negative BOLD affected many areas connected to the ascending reticular activating system (ARAS). The ARAS was decoupled to much of the brain but was hyperconnected to the olfactory system and prefrontal cortex.ConclusionThe CBD-induced decrease in ARAS activity is consistent with an emerging literature suggesting that CBD reduces autonomic arousal under conditions of emotional and physical stress.

Bayesian Calibration of a Computational Model of Tissue Expansion Based on a Porcine Animal Model

Tissue expansion is a technique used clinically to grow skin in situ to correct large defects. Despite its enormous potential, the use of tissue expansion is limited by our inability to control skin growth at will. We lack fundamental knowledge of how skin adapts to mechanical cues, and we lack computational models to predict growth to guide clinical treatment. In our previous work, we introduced a finite element model of tissue expansion that predicted key patterns of strain and growth which were then confirmed by a porcine animal model also introduced by us. Here we use the data from a new set of experiments on the animal model to calibrate the computational model in a Bayesian framework. We find that growth can be explained based on the total deformation. The characteristic area growth rate is k e [0.01,0.022] [1/hrs]. Growth is anisotropic, the rostral-caudal axis shows greater deformation than the transverse axis, and the time scale of growth in the rostral-caudal direction is given by a rate parameters k1 e in the range [0.01,0.035 ] [1/hrs] compared to k2 e of [0.003,0.008] [1/hrs] in the transverse direction. Moreover, the results underscore the high variability in biological systems, and the need to create probabilistic computational models to predict tissue adaptation in realistic settings

Mapping the orbitofrontal cortex using temporal fluctuations in cerebral blood flow

IntroductionThe orbitofrontal cortex (OFC) is involved in diverse cognitive and behavioral processes including incentive valuation, decision‐making, and reinforcement learning. Anatomic and cytoarchitectonic studies divide the OFC along both medial‐lateral and rostral‐caudal axes. OFC regions diverge in structure and function, assessed in vivo using white matter tractography and blood oxygenation level‐dependent (BOLD) MRI, respectively. However, interpretation of T 2*‐weighted BOLD is limited by susceptibility artifacts in the inferior frontal lobes, with the spatial pattern of these artifacts frequently assuming the geometry of OFC organization. Here, we utilize a novel perfusion‐weighted arterial spin labeling (ASL) functional connectivity approach, which is minimally susceptibility‐weighted, to test the hypothesis that OFC topology reflects correlated temporal hemodynamic activity.MethodsIn healthy participants (n = 20; age = 29.5 ± 7.3), 3D ASL scans were acquired (TR/TE = 3,900/13 ms; spatial resolution = 3.8 mm isotropic). To evaluate reproducibility, follow‐up scanning on a separate day was performed on a participant subset (n = 8). ASL‐based connectivity was modeled for gray matter OFC voxels, and k‐means clustering (k = 2–8) applied to correlation statistics.ResultsThese approaches revealed both medial‐lateral and rostral‐caudal OFC divisions, confirming our hypothesis. Longitudinal reproducibility testing revealed 84% voxel clustering agreement between sessions for the k = 2 solution.ConclusionTo our knowledge, this constitutes the first in vivo cortical parcellation based on perfusion fluctuations. Our approach confirms functional OFC subdivisions predicted from anatomy using a less susceptibility‐sensitive method than the conventional approach.

Gradients of functional organization in posterior parietal cortex revealed by visual attention, visual short-term memory, and intrinsic functional connectivity

Visual attention and visual working memory tasks recruit a common network of lateral frontal cortical (LFC) and posterior parietal cortical (PPC) regions. Here, we examine finer-scale organization of this frontoparietal network. Three LFC regions recruited by visual cognition tasks, superior precentral sulcus (sPCS), inferior precentral sulcus (iPCS), and mid inferior frontal sulcus (midIFS) exhibit differential patterns of resting-state functional connectivity to PPC. A broad dorsomedial to ventrolateral gradient is observed, with sPCS connectivity dominating in the dorsomedial PPC band, iPCS dominating in the middle band, and midIFS dominating in the ventrolateral band. These connectivity-defined subregions of PPC capture differential task activation between a pair of visual attention and working memory tasks. The relative functional connectivity of sPCS and iPCS also varies along the rostral-caudal axis of the retinotopic regions of PPC. iPCS connectivity is relatively stronger near the IPS0/IPS1 and IPS2/IPS3 borders, especially on the lateral portions of these borders, which each preferentially encode central visual field representations. In contrast, sPCS connectivity is relatively stronger elsewhere in retinotopic IPS regions which preferentially encode peripheral visual field representations. These findings reveal fine-scale gradients in functional connectivity within the frontoparietal visual network that capture a high-degree of specificity in PPC functional organization.

Double UP: A Dual Color, Internally Controlled Platform for in utero Knockdown or Overexpression.

In utero electroporation (IUE) is a powerful tool for testing the role of genes in neuronal migration and function, but this technique suffers from high degrees of variability. Such variability can result from inconsistent surgery, developmental gradients along both rostral-caudal and medial-lateral axes, differences within littermates and from one litter to another. Comparisons between control and experimental electroporations rely on section matching, which is inherently subjective. These sources of variability are cumulative, leading to difficult to interpret data and an increased risk of both false positives and false negatives. To address these limitations, we developed two tools: (1) a new plasmid, termed Double UP, which combines LoxP-flanked reporters and limiting Cre dosages to generate internal controls, and (2) an automated program for unbiased and precise quantification of migration. In concert, these tools allow for more rigorous and objective experiments, while decreasing the mice, time, and reagents required to complete studies.

Invited Commentary on "Optimizing Diffusion-Tensor Imaging Acquisition for Spinal Cord Assessment," with Response from Dr Martín Noguerol et al.

Invited Commentary on "Optimizing Diffusion-Tensor Imaging Acquisition for Spinal Cord Assessment," with Response from Dr Martín Noguerol et al.

Development of the brainstem respiratory circuit.

The respiratory circuit is comprised of over a dozen functionally and anatomically segregated brainstem nuclei that work together to control respiratory rhythms. These respiratory rhythms emerge prenatally but only acquire vital importance at birth, which is the first time the respiratory circuit faces the sole responsibility for O2 /CO2 homeostasis. Hence, the respiratory circuit has little room for trial-and-error-dependent fine tuning and relies on a detailed genetic blueprint for development. This blueprint is provided by transcription factors that have specific spatiotemporal expression patterns along the rostral-caudal or dorsal-ventral axis of the developing brainstem, in proliferating precursor cells and postmitotic neurons. Studying these transcription factors in mice has provided key insights into the functional segregation of respiratory control and the vital importance of specific respiratory nuclei. Many studies converge on just two respiratory nuclei that each have rhythmogenic properties during the prenatal period: the preBötzinger complex (preBötC) and retrotrapezoid nucleus/parafacial nucleus (RTN/pF). Here, we discuss the transcriptional regulation that guides the development of these nuclei. We also summarize evidence showing that normal preBötC development is necessary for neonatal survival, and that neither the preBötC nor the RTN/pF alone is sufficient to sustain normal postnatal respiratory rhythms. Last, we highlight several studies that use intersectional genetics to assess the necessity of transcription factors only in subregions of their expression domain. These studies independently demonstrate that lack of RTN/pF neurons weakens the respiratory circuit, yet these neurons are not necessary for neonatal survival because developmentally related populations can compensate for abnormal RTN/pF function at birth. This article is categorized under: Nervous System Development > Vertebrates: Regional Development.

Wnt-Notch Signaling Interactions During Neural and Astroglial Patterning of Human Stem Cells.

The human brain formation involves complicated processing, which is regulated by a gene regulatory network influenced by different signaling pathways. The cross-regulatory interactions between elements of different pathways affect the process of cell fate assignment during neural and astroglial tissue patterning. In this study, the interactions between Wnt and Notch pathways, the two major pathways that influence neural and astroglial differentiation of human induced pluripotent stem cells (hiPSCs) individually, were investigated. In particular, the synergistic effects of Wnt-Notch pathway on the neural patterning processes along the anterior-posterior or dorsal-ventral axis of hiPSC-derived cortical spheroids were explored. The human cortical spheroids derived from hiPSCs were treated with Wnt activator CHIR99021 (CHIR), Wnt inhibitor IWP4, and Notch inhibitor (N-[N-(3,5-difluorophenacetyl)-l-alanyl]-S-phenylglycine t-butyl ester [DAPT]) individually, or in combinations (CHIR + DAPT, IWP4 + DAPT). The results suggest that CHIR + DAPT can promote Notch signaling, similar or higher than CHIR alone, whereas IWP4 + DAPT reduces Notch activity compared to IWP4 alone. Also, CHIR + DAPT promoted hindbrain marker HOXB4 expression more consistently than CHIR alone, while IWP4 + DAPT promoted Olig2 expression, indicating the synergistic effects distinctly different from that of the individual small molecule. In addition, IWP4 simultaneously promoted dorsal and ventral identity. The patterned neural spheroids can be switched for astroglial differentiation using bone morphogenetic protein 4. This study should advance the derivations of neurons, astroglial cells, and brain region-specific organoids from hiPSCs for disease modeling, drug screening, as well as for hiPSC-based therapies. Impact Statement Wnt signaling plays a central role in neural patterning of human pluripotent stem cells. It can interact with Notch signaling in defining dorsal-ventral and rostral-caudal (or anterior-posterior) axis of brain organoids. This study investigates novel Wnt and Notch interactions (i.e., Wntch) in neural patterning of dorsal forebrain spheroids or organoids derived from human induced pluripotent stem cells. The synergistic effects of Wnt activator or inhibitor with Notch inhibitor were observed. This study should advance the derivations of neurons, astroglial cells, and brain region-specific organoids from human stem cells for disease modeling and drug screening, as well as for stem cell-based therapies. The results can be used to establish better in vitro culture methods for efficiently mimicking in vivo structure of central nervous system.

Global Jitter Motion of the Retinal Image Dynamically Alters the Receptive Field Properties of Retinal Ganglion Cells.

Fixational eye movements induce aperiodic motion of the retinal image. However, it is not yet fully understood how fixational eye movements affect retinal information processing. Here we show that global jitter motion, simulating the image motion during fixation, alters the spatiotemporal receptive field properties of retinal ganglion cells. Using multi-electrode and whole-cell recording techniques, we investigated light-evoked responses from ganglion cells in the isolated goldfish retina. Ganglion cells were classified into six groups based on the filtering property of light stimulus, the membrane properties, and the cell morphology. The spatiotemporal receptive field profiles of retinal ganglion cells were estimated by the reverse correlation method, where the dense noise stimulus was applied on the dark or random-dot background. We found that the jitter motion of the random-dot background elongated the receptive filed along the rostral-caudal axis and temporally sensitized in a specific group of ganglion cells: Fast-transient ganglion cells. At the newly emerged regions of the receptive field local light stimulation evoked excitatory postsynaptic currents with large amplitude and fast kinetics without changing the properties of inhibitory postsynaptic currents. Pharmacological experiments suggested two presynaptic mechanisms underlying the receptive field alteration: (i) electrical coupling between bipolar cells, which expands the receptive field in all directions; (ii) GABAergic presynaptic inhibition from amacrine cells, which reduces the dorsal and ventral regions of the expanded receptive field, resulting in elongation along the rostral-caudal axis. Our study demonstrates that the receptive field of Fast-transient ganglion cells is not static but dynamically altered depending on the visual inputs. The receptive field elongation during fixational eye movements may contribute to prompt firing to a target in the succeeding saccade.

MAP2 immunoreactivity deficit is conserved across the cerebral cortex within individuals with schizophrenia

Several postmortem studies have reported lower levels of immunoreactivity (IR) for microtubule-associated protein 2 (MAP2) in several cortical regions of individuals with schizophrenia (SZ). However, whether this effect is conserved across multiple brain areas within an individual with SZ or if it is regionally-specific remains unclear. We characterized patterns of MAP2-IR across three cortical regions at different levels of the rostral-caudal axis within individual subjects with and without SZ. MAP2-IR levels were measured in deep layer 3 of dorsolateral prefrontal cortex (DLPFC), lateral intraparietal cortex (LIP), and primary visual cortex (V1). Postmortem tissue containing each cortical region was derived from 20 pairs of SZ subjects and nonpsychiatric comparison (NPC) subjects matched perfectly for sex, and as closely as possible for age and postmortem interval. MAP2-IR was assessed by quantitative fluorescence microscopy. We observed significantly lower levels of MAP2-IR in SZ subjects relative to NPC subjects, without a significant region by diagnosis interaction. Logs of the within-pair ratios (SZ:NPC) of MAP2-IR were significantly correlated across the three regions. These findings demonstrate that MAP2-IR deficits in SZ are consistent across three neocortical regions within individual subjects. This pattern of MAP2-IR deficit has implications for therapeutic development and future investigations of MAP2 pathology in SZ.

A tale of two tracts: history, current advances, and future directions of research on sexual differentiation of reproductive tracts†.

Alfred Jost's work in the 1940s laid the foundation of the current paradigm of sexual differentiation of reproductive tracts, which contends that testicular hormones drive the male patterning of reproductive tract system whereas the female phenotype arises by default. Once established, the sex-specific reproductive tracts undergo morphogenesis, giving rise to anatomically and functionally distinct tubular organs along the rostral-caudal axis. Impairment of sexual differentiation of reproductive tracts by genetic alteration and environmental exposure are the main causes of disorders of sex development, and infertility at adulthood. This review covers past and present work on sexual differentiation and morphogenesis of reproductive tracts, associated human disorders, and emerging technologies that have made impacts or could radically expand our knowledge in this field.

Cux2 refines the forelimb field by controlling expression of Raldh2 and Hox genes.

ABSTRACTIn vertebrates, two pairs of buds that give rise to the fore- and hindlimbs form at discrete positions along the rostral-caudal axis of the body. The mechanism responsible for the positioning of the limb buds is still largely unknown. Here we show a novel function for Cut homeobox transcription factor 2 (Cux2), the ortholog of Drosophila cut, in refining the forelimb field during chick development. Cux2 is expressed in the forelimb field before the emergence of the limb buds. Knocking down the expression of Cux2 using small interfering RNA (siRNA) resulted in a caudal shift of the forelimb bud, whereas misexpression of Cux2 or the constitutively active Cux2-VP16 caused a rostral shift of the forelimb bud or reduction of the forelimb field along the anterior-posterior axis. Further functional analyses revealed that expression of Hoxb genes and retinaldehyde dehydrogenase 2 (Raldh2), which are involved in limb positioning, are directly activated by Cux2 in the lateral plate mesoderm. Our data suggest that Cux2 in the lateral plate mesoderm refines the forelimb field via regulation of Raldh2 and Hoxb genes in chicken embryos.

Antagonism between the transcription factors NANOG and OTX2 specifies rostral or caudal cell fate during neural patterning transition

During neurogenesis, neural patterning is a critical step during which neural progenitor cells differentiate into neurons with distinct functions. However, the molecular determinants that regulate neural patterning remain poorly understood. Here we optimized the "dual SMAD inhibition" method to specifically promote differentiation of human pluripotent stem cells (hPSCs) into forebrain and hindbrain neural progenitor cells along the rostral-caudal axis. We report that neural patterning determination occurs at the very early stage in this differentiation. Undifferentiated hPSCs expressed basal levels of the transcription factor orthodenticle homeobox 2 (OTX2) that dominantly drove hPSCs into the "default" rostral fate at the beginning of differentiation. Inhibition of glycogen synthase kinase 3β (GSK3β) through CHIR99021 application sustained transient expression of the transcription factor NANOG at early differentiation stages through Wnt signaling. Wnt signaling and NANOG antagonized OTX2 and, in the later stages of differentiation, switched the default rostral cell fate to the caudal one. Our findings have uncovered a mutual antagonism between NANOG and OTX2 underlying cell fate decisions during neural patterning, critical for the regulation of early neural development in humans.

Self-patterning of rostral-caudal neuroectoderm requires dual role of Fgf signaling for localized Wnt antagonism

The neuroectoderm is patterned along a rostral-caudal axis in response to localized factors in the embryo, but exactly how these factors act as positional information for this patterning is not yet fully understood. Here, using the self-organizing properties of mouse embryonic stem cell (ESC), we report that ESC-derived neuroectoderm self-generates a Six3+ rostral and a Irx3+ caudal bipolarized patterning. In this instance, localized Fgf signaling performs dual roles, as it regulates Six3+ rostral polarization at an earlier stage and promotes Wnt signaling at a later stage. The Wnt signaling components are differentially expressed in the polarized tissues, leading to genome-wide Irx3+ caudal-polarization signals. Surprisingly, differentially expressed Wnt agonists and antagonists have essential roles in orchestrating the formation of a balanced rostral-caudal neuroectoderm pattern. Together, our findings provide key processes for dynamic self-patterning and evidence that a temporally and locally regulated interaction between Fgf and Wnt signaling controls self-patterning in ESC-derived neuroectoderm.

Vitamin A induced homeotic hindlimb formation on dorsal and ventral sides of regenerating tissue of amputated tails of Japanese brown frog tadpoles.

When anuran tadpoles are treated with vitamin A after tail amputation, hindlimb-like structures can be generated instead of the lost tail part at the amputation site. This homeotic transformation was initially expected to be a key to understanding the body plan of vertebrates. Unfortunately, homeotic limb formation has been reproduced in only some Indian frog species and a European species, but not in experimental anurans such as Xenopus laevis or Rana catesbeiana. Consequently, this fascinating phenomenon has not been well analyzed, especially at the molecular level. In addition, the initial processes of ectopic limb development are also unclear because morphological changes in the early phases have not been analyzed in detail. In this study, we report the induction of homeotic transformation using Japanese brown frogs and present a detailed morphological analysis. Unexpectedly, the ectopic limbs developed not only at the ventral sites, but also at the dorsal sites of the tail regenerates of vitamin A-treated tadpoles. The relationship between position and axial orientation of ectopic limbs suggested the double duplication of positional value order along the rostral-caudal axis and the dorsal-ventral axis of the tail regenerates.

Links Between Patterns of Embryonic Spinal Cord Development and Schizophrenia Risk

The entry point of the spermatozoon dictates the placement of the animal pole and thus the dorsoventral axis of the early embryo, eliminating the once-radial symmetry of the egg. Radial symmetry is further broken by the primitive streak, demarcating the left–right and rostral–caudal axes. This newfound bilateral symmetry is short-lived, however, disturbed by development of the (generally) right-sided loop formation of the heart tube and subsequent asymmetrically placed organ systems, such as the liver, spleen, and digestive tract ( 1 Levin M. Left-right asymmetry in embryonic development: A comprehensive review. Mech Dev. 2005; 122: 3-25 Crossref PubMed Scopus (373) Google Scholar ). Left–Right Asymmetry of Maturation Rates in Human Embryonic Neural DevelopmentBiological PsychiatryVol. 82Issue 3PreviewLeft–right asymmetry is a fundamental organizing feature of the human brain, and neuropsychiatric disorders such as schizophrenia sometimes involve alterations of brain asymmetry. As early as 8 weeks postconception, the majority of human fetuses move their right arms more than their left arms, but because nerve fiber tracts are still descending from the forebrain at this stage, spinal–muscular asymmetries are likely to play an important developmental role. Full-Text PDF

Orthogonal topography in the parallel input architecture of songbird HVC

Neural activity within the cortical premotor nucleus HVC (acronym is name) encodes the learned songs of adult male zebra finches (Taeniopygia guttata). HVC activity is driven and/or modulated by a group of five afferent nuclei (the Medial Magnocellular nucleus of the Anterior Nidopallium, MMAN; Nucleus Interface, NIf; nucleus Avalanche, Av; the Robust nucleus of the Arcopallium, RA; the Uvaeform nucleus, Uva). While earlier evidence suggested that HVC receives a uniformly distributed and nontopographic pattern of afferent input, recent evidence suggests this view is incorrect (Basista et al., ). Here, we used a double-labeling strategy (varying both the distance between and the axial orientation of dual tracer injections into HVC) to reveal a massively parallel and in some cases topographic pattern of afferent input. Afferent neurons target only one rostral or caudal location within medial or lateral HVC, and each HVC location receives convergent input from each afferent nucleus in parallel. Quantifying the distributions of single-labeled cells revealed an orthogonal topography in the organization of afferent input from MMAN and NIf, two cortical nuclei necessary for song learning. MMAN input is organized across the lateral-medial axis whereas NIf input is organized across the rostral-caudal axis. To the extent that HVC activity is influenced by afferent input during the learning, perception, or production of song, functional models of HVC activity may need revision to account for the parallel input architecture of HVC, along with the orthogonal input topography of MMAN and NIf.